FR2890859A1 - Composition useful as a tyrosinase expression inhibitor e.g. to depigment the skin, comprises at least one double stranded RNA oligonucleotide associated to a cationic particle having a specified size and zeta potential - Google Patents

Abstract

Composition comprises at least one double stranded RNA oligonucleotide (I) associated with a cationic particle of =1 mu m in size and 10-80 mV of zeta potential. Composition comprises at least one double stranded RNA oligonucleotide (I) (SEQ ID No. 1-48 comprising one of 48 fully defined 19-25 amino acid sequences given in the specification) associated with a cationic particle of =1 mu m in size and 10-80 mV of zeta potential. An independent claim is included for cosmetic process to bleach, clear up the color and/or uniform the color of a browned skin comprising topical application of (I). ACTIVITY : Dermatological. MECHANISM OF ACTION : Tyrosinase expression inhibitor. The ability of (I) on the degradation of tyrosinase messenger RNA was tested. The results showed that the double stranded RNA oligonucleotide of SEQ ID No. 43 comprising a fully defined 25 amino acid sequences given in the specification, exhibited 99.1% inhibition of tyrosinase.

Description

The present invention relates to novel RNA oligonucleotides

  double strand for decreasing the expression of tyrosinase, their use for cosmetic and / or pharmaceutical purposes as well as their association with cationic particles of less than or equal to 1 μm, zeta potential between 10 and 80 mV.

  Tyrosinase (monophenol dihydroxyl phenylalanine: oxygen oxidoreductase EC 1.14.18.1) is the essential enzyme involved in this series of reactions. In particular, it catalyzes the conversion reaction of tyrosine to Dopa (dihydroxyphenylalanine) thanks to its hydroxylase activity and the conversion reaction of Dopa to dopaquinone by virtue of its oxidase activity. This tyrosinase only acts when it is maturing under the influence of certain biological factors.

  This enzyme may also be of interest in the treatment of pathologies such as melanoma (Riley et al., J. Immunother, 2001, 21, 212-220) or Vogt-Koyanagi-Harada disease (Read et al., Ophthalmol., 2000, 11, 437-442).

  The mechanism of formation of skin pigmentation, that is to say the formation of melanin is particularly complex and involves schematically the following main steps: Tyrosine -> Dopa - * Dopaquinone - * Dopachrome * Melanin Thus The pigmentation of human skin results from the synthesis of melanin by dendritic cells, melanocytes. These contain organelles called melanosomes, seat of melanin biosynthesis. These are melanosomes which, after migration along the dendrites, are transferred from melanocytes to keratinocytes. Keratinocytes are then transported to the surface of the skin during the process of differentiation of the epidermis (Gilchrest BA, Park HY, Eller MS, Yaar M, Mechanisms of ultraviolet light-induced pigmentation, Photochem Photobiol 1996, 63: 1 10, Hearing VJ, Tsukamoto K, Enzymatic control of pigmentation in mammals, FASEB J 1991, 5: 2902-2909).

  Among the enzymes of melanogenesis, tyrosinase is a key enzyme that catalyzes the first two steps of melanin synthesis. Homozygous tyrosinase mutations cause type 1 oculocutaneous albinism characterized by a complete absence of melanin synthesis (Toyofuku K, Wada Spritz RA, Hearing VJ, The molecular basis of oculocutaneous albinism type 1 (OCA1)). mutant tyrosinase results in lack of pigmentation Biochem J. 2001; 355: 259-269).

  The hyperpigmentation disorders resulting from an increase in melanin production, new therapeutic approaches whose rational is based on the inhibition of tyrosinase activity are important to develop.

  Most skin lightening compounds already known are phenols / catechols.

  These compounds inhibit tyrosinase but the majority of these compounds are cytotoxic towards melanocytes, which could cause permanent depigmentation of the skin.

  It therefore seems interesting, for an application in humans, to have new tyrosinase inhibiting compounds having both good efficacy and good tolerance.

  Recently, the use of oligonucleotides of double-stranded RNA, dsRNA and more particularly of siRNA (from 12 to 40 nucleotides), would make it possible to obtain a specific activity in the cosmetic field such as skincare or skincare. capillary, but also in the dermatological and pharmaceutical fields.

  However, the in vivo use of siRNAs is known to present various difficulties. In addition to the problem of the penetration of these siRNAs to reach the target cells during their topical application, particularly because of the difficulty in crossing the stratum corneum, experience has shown that the administration of siRNA can lead to the triggering of an answer. interferon reported by numerous publications (Sledz CA et al., Activation of the interferon system by short-interfering RNAs Nat Cell Biol., 2003; 9: 834-9. Katalin Karikô et al., Small Interfering RNAs Sequence-Independent Gene Mediate Suppression and Induction Immune Activation by Signaling through Toll-Like Receptor 31. J. Immunol 2004; 172: 6545-6549 Judge AD et al., Sequence-dependent stimulation of the mammalian innate immune response by synthetic siRNA, Nat Biotechnol 2005; 23: 457-62) as well as the induction or inhibition of non-target gene expression by siRNA (Jackson et al., Expression profiling reveals off-target gene regulation by RNA / Nat Biotechnol 2003). ; 21: 635 -7), these two phenomena are highly undesirable.

  US patent application 2004/0215006 discloses anti-sense and double-stranded single stranded RNA oligonucleotides active against tyrosinase, the examples used relate solely to antisense RNA single-stranded oligonucleotides. In particular, siRNAs homologous to these antisense RNAs have not been shown to be effective.

  Some authors have pointed out that double-stranded RNA oligonucleotides, such as siRNAs, and single-strand antisense RNA oligonucleotides had different targets and that the activity of an antisense RNA was not extrapolable to siRNA of the same sequence (Xu et al., Effective small interfering RNAs and phosphorothioate antisense DNAs have different preferences for mRNAs luciferase sites.) BBRC 2003; 306: 712717).

  Furthermore, in this document US 2004/0215006, the proposed siRNA concentrations are between 50 and 200 nM. As noted above, it has been described by Jackson et al. strong positive and negative regulation of non-targeted siRNA genes at concentrations of 100 nM.

  Similarly, in WO 2005/060536 which describes siRNAs specifically inhibiting tyrosinase, the concentration ranges proposed are very broad and can lead to compositions comprising a very large quantity of siRNAs for which it will be possible to observe a positive regulation and negative for non-siRNA targeted genes. In the context of a cosmetic or therapeutic use, this regulation of non-targeted genes is not acceptable, particularly because of its unpredictability.

  The Applicant has developed new siRNA specifically inhibiting the expression of tyrosinase from which it has selected siRNAs having an activity such that these siRNAs can be used at a dose of less than or equal to 1 nM.

  In addition to solving the problems related to the induction of nonspecific genes, the applicant verified that these sequences did not induce an interferon response.

  The Applicant verified that the selected siRNAs did not induce an interferon type response. For this, it measured the expression of the OAS-1 and IFIT-1 (interferon-inducible tetratricopeptide repeat domain) genes known to be induced by interferon (Stefan F. Wieland et al., Searching for Interferon-Induced Genes That Inhibit Hepatitis B Virus Replication in Transgenic Mouse Hepatocytes, J. of Virology 2003, 77: 12271236) according to the protocol described in the kit manual BLOCK-iTTM RNAi Stress Response Control Kit (Human) for interferon-mediated stress response to dual-response stranded RNA in human cells marketed by Invitrogen.

  The combination of these tyrosinase-specific siRNAs with cationic particles having a size of less than or equal to 1 μm and a zeta potential of between 10 and 80 mV makes it possible to very significantly improve their penetration into the target cells of a model. three-dimensional as the skin. Once penetrated, the tyrosinase-specific siRNA becomes active.

  Penetration can be assayed using siRNA-fixed fluorescent marker and its activity by quantitating the targeted messenger by quantitative PCR or by assaying the corresponding target messenger protein.

  The cationic particles of the invention may be surfactant micelles, micelles of block or non-block polymers, cationic liposomes and niosomes, cationic oleosomes, cationic nanoemulsions, as well as cationic organic or inorganic particles and nanocapsules.

  Thus, a first subject of the invention relates to a double-stranded RNA oligonucleotide (also called dsRNA or siRNA) chosen from oligonucleotides of sequence SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 and 48, optionally modified.

  The use of dsRNA and, more particularly, siRNA (for short interfering RNA) makes it possible to obtain a specific inhibition activity for the synthesis of a target protein by degradation of the mRNA encoding the protein. The degradation of the target mRNA is achieved by the activation of the effective RISC (RNA Induced Silencing Complex) by the attachment of the dsRNA antisense strand to the mRNA (see Tuschl T. Chem Biochem 2001; 239-245, Nykanen A et al, Cell 2001, 107: 309-321, Dorsett Y. and Tuschl T. Nat Rev Drug Discov 2004, 3: 318-329, Downward J. BMJ 2004, 328: 1245-1248; Shankar P. et al., JAMA 2005; 293: 1367-1373).

  The molecular mechanism involved involves double-stranded RNA fragments consisting of 12 to 40 nucleotides, preferably 23 to 27 nucleotides.

  These double-stranded RNA oligonucleotides may preferentially be composed of a homologous sense strand and an antisense strand complementary to the sequence of the mRNA of human tyrosinase (GenBank accession number NM_000372). The double-stranded RNA oligonucleotide has blunt or unpaired ends of 2 to 6 nucleotides.

  The double-stranded RNA oligonucleotides can be synthesized according to numerous in vivo or in vitro synthesis methods manually or automatically.

  In vitro synthesis methods can be chemical or enzymatic, for example by using an RNA polymerase (as an example of T3, T7 or SP6) which will carry out the transcription of a DNA sequence template (or cDNA ) chosen.

  Many methods of in vivo synthesis of double-stranded RNA are described in the literature, they can be performed in various cell types of bacteria or higher organisms (Sambrook et al., Molecular Cloning, A Laboratory Manual, Second Edition (1989)). ), DNA cloning, volume I and II, DN Glover (ed., 1985), Oligonucleotide Synthesis, MJ Gaits (ed., 1984), Nucleic Acid Hybridization, BD Hames and SJ Higgins (ed.1984), Transcription and Translation BD Hames SJ Higgins (ed.1984), Animal Cell Culture, RI Freshney (1986), Immobilized Cells and Enzymes, IRL Press (1986), B. Pertal, A Practical Guide to Molecular Cloning (1984), Gene Transfer Vectors for Mammalian Cells, JH Miller and MP Ca / os, Cold Spring Harbor Laboratory (eds. 1987), Methods in Enzymology, vol.154, Wu and Grossman, and 155, Wu, Mayer and Walker (1987), Immunochemical Methods in Cell and Molecular. Biology, Academic Press, London, Scopes (1987), Protein Purificatio n: Principle and Practice, 2nd ed., Springer-Verlag, N.-Y. and Handbook of Experimental Immunology, vol. IV, C. D. Weir and C. C. Blackwell (1986)). Reference may also be made to the methods of synthesis described in patent applications W001 / 36646, W001 / 75164 and US20030088087.

  Preferably, the double-stranded RNA oligonucleotides according to the invention are modified by the addition to 2 'of a -O-Methyl group.

  The oligonucleotides of double-stranded RNA can undergo various modifications, they can in particular be modified as described in document US 2004014956.

  Advantageously, the modifications will lead to more stable double-stranded RNA oligonucleotides and, more preferably, double-stranded RNA oligonucleotides rendered stealthy with respect to the interferon response, this property is crucial for in vivo use. . Double-stranded RNA oligonucleotides may still have a modified sense sequence so that it can not be incorporated into the RISC and therefore can not induce side effects.

  Preferably, the double-stranded RNA oligonucleotides of SEQ will be used. ID. Nos. 1, 2, 4, 13, 16, 35, 37, 40 and 42 having a degradation efficiency of tyrosinase mRNA greater than 50% at 0.016 nM measured according to the protocol of Example 2 below.

  The subject of the present invention also relates to a composition comprising at least one double-stranded RNA oligonucleotide chosen from oligonucleotides of sequence SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 and 48.

  In this composition, the double-stranded RNA oligonucleotide is present at a concentration of less than or equal to 100 μM, preferably less than or equal to 10 μM and still more preferably at 1 μM.

  The composition according to the invention is preferably adapted for topical administration on the body surface, that is to say, the skin, superficial body growths, mucous membranes and eyes.

  In a preferred embodiment of the present invention, the double-stranded RNA oligonucleotides are associated with a cationic particle having a size of less than or equal to 1 μm and a zeta potential of between 10 and 80 mV.

  In this embodiment, the double-stranded RNA oligonucleotides will adhere to the surface of the particle, which serves as a vehicle for penetrating the cutaneous structures and in the target cells of the skin, mucous membranes or even mucous membranes. eyes for cosmetic, dermatological and / or ophthalmic applications.

  The cationic particles according to the invention are particles having a size less than or equal to 1 μm, preferably less than or equal to 500 nm, more preferably less than or equal to 300 nm, measurable with, for example, a laser particle size analyzer type BI90 plus from the company Brookhaven, and having a zeta potential between 10 and 80 mV can be measured with a Coultronics DELSA 440 type zetameter.

  The cationic particle may be chosen from surfactant micelles, in particular micelles of nonionic amphiphilic surfactants and cationic surfactants, micelles of block polymers, in particular cationic amphiphilic block polymer micelles, amphiphilic block polymer micelles. nonionic and cationic amphiphilic block polymer and micelles of nonionic amphiphilic block polymer and cationic surfactant, liposomes of nonionic and cationic surfactants, niosomes, oleosomes, nanoemulsion particles, nanocapsules, organic particles or inorganic particles.

  Listed below and in a non-exhaustive manner are cationic particles that can be used according to the invention.

  Micelles of surfactants As a reminder, micelles are aggregates that spontaneously form amphiphilic molecules when they are solubilized in water or oil, beyond a certain so-called critical concentration: CMC.

  The micelles that can be used in the context of the invention consist of at least one cationic surfactant. This cationic surfactant may be combined with one or more nonionic amphiphilic surfactants.

  Those skilled in the art will advantageously select the nonionic and cationic surfactants in the McCutcheons 1998 emulsifiers and detergents as well as in the following editions.

  As non-limiting examples, the cationic surfactants that can be used in the context of the invention are listed below.

  Without being limiting, the nonionic surfactants that can be used are: alkyls and polyalkyls (C6 to C30, saturated or unsaturated, branched or unbranched), ester or ether of POE, glycerol and polyglycerol, oxyethylenated sorbitan or otherwise, sucrose , glucose oxyethylenated or not, maltose, POP-POE.

  In case of mixing between nonionic surfactants and cationic surfactants, their respective proportions, by weight, will be between 99/1 and 1/99.

  The level of surfactants forming the micelles will be dependent on the CMC thereof. However, in the context of the invention, the concentration of micellar surfactants will be between 0.1 and 10% and preferably between 0.2 and 5% by weight relative to the total weight of the composition.

  Block Polymer Micelles Amiphilic block polymer micelles may be prepared according to the method described in WO 04/035013.

  The block copolymers that are useful for the preparation of the micelles associated with the dsRNAs according to the invention are, in particular, amphiphilic block polymers, preferably nonionic, diblock or triblock polymers, which can form micelles in contact with water. They are in particular diblock type (A-B) or triblock (A-B-A), A corresponding to a nonionic hydrophilic polymeric block and B to a hydrophobic polymeric block. The molecular weight of the polymers can be between 1000 and 100 000 and the ratio A / B can be between 1/100 and 50/1.

  In the context of the invention, three types of micelles can be used: micelles of cationic amphiphilic block polymer; mixed micelles of nonionic amphiphilic block polymer associated with a cationic amphiphilic block polymer; and mixed micelles of nonionic amphiphilic block polymer associated with a cationic surfactant.

  The nonionic hydrophilic polymer block may be chosen from polyethylene oxide (POE) and polyvinylpyrrolidone (PVP).

  The hydrophobic polymeric block may be chosen from polystyrene, poly (tert.-butylstyrene), poly (methylmethacrylate), poly (ethylacrylate), poly (butylacrylate), poly (butylmethacrylate), poly (vinylacetate), polycaprolactones, polycaprolactams, polydimethylsiloxanes, C3-C6 alkylene polyoxides, polyaspartic acid, polylactic acid, poly glycolic acid, polyleucine, polybutadienes, polyethylenes, polypropylenes, polybutylenes.

  The block copolymer is preferably chosen from the following block copolymers: propylene polyoxide / ethylene oxide polystyrene / polyoxyethylene / polymethylmethacrylate / polyoxyethylene polybutylmethacrylate / polyoxyethylene / polyoxybutylene / polyoxyethylene polycaprolactone / polyoxyethylene / polyethylene / polyoxyethylene polyoxyethylene / polyoxybutylene / polyoxyethylene.

  In the context of the invention, it is necessary to add to the micellar composition: a cationic amphiphilic block polymer of which one of the blocks is cationic and may be chosen from, for example: polytrimethyl methacrylate ethyl ammonium; Dimethylaminoethyl quaternized polymethylmethacrylate; Poly methyl vinyl imidazolium; Poly vinyl benzyl trimethylammonium chloride.

  the combination of a nonionic amphiphilic block polymer with a cationic amphiphilic block polymer is such that the ratio between the two will be between 99/1 and 1/99; and / or at least one cationic surfactant as listed below.

  In this case, the respective ratio between the nonionic amphiphilic block polymer and the cationic surfactant will be between 50/50 and 99/1.

  In the context of the invention, the concentration of micellar block polymers associated or not with a cationic surfactant, will be between 0.1 and 10% and, preferably, between 0.2 and 5% by weight relative to the total weight of the composition.

  In one variant of the invention, it is also possible to form micelles constituted by cationic amphiphilic block polymers as described above.

  Liposomes and Niosomes The nonionic amphiphilic lipids capable of forming nonionic liposomes are in particular those described in the patent application EP 0 582 503.

  In particular, the nonionic amphiphilic lipids may consist of a mixture of esters of at least one polyol selected from the group consisting of polyethylene glycol having 1 to 60 ethylene oxide units, sorbitan, sorbitan carrying 2 60 units of ethylene oxide, glycerol carrying 2 to 30 ethylene oxide units, polyglycerols comprising 2 to 15 glycerol units, sucroses, glucose containing 2 to 30 ethylene oxide units, and at least one fatty acid having a C5-C17 alkyl chain, saturated or unsaturated, linear or branched, the number of alkyl chains per polyol group being between 1 and 10.

  The term "ester mixture" not only covers mixtures of pure esters of different chemical families but also covers any product which contains several chemically pure polyol esters of the same family in varying proportions. It is, in particular, the case of the products having a statistical formula, in their hydrophilic part, for example a polyglycerol ester of formula CO- (OCH 2 -CHOH-CH 2) n-OH where n is a statistical value and which can contain various proportions of esters for which n = 1, n = 2, n = 3, n = 4, etc ...; this is also the case for esters containing several alkyl chains in their lipophilic part, such as cocoates, which contain C5 to C17 alkyl chains or isostearates, where the C17 alkyl chains are a complex mixture of isomeric forms; this is also the case for products consisting of mixtures of mono-, di-, tri- or polyesters of the same polyol. It should be noted that a product which contains only one ester capable of forming vesicles and impurities of another type could not be used according to the invention.

  Commercial esters which can be used alone according to the invention, since they are in reality mixtures of esters, are for example the following: partial esters of sorbitan (or sorbitol anhydride) and of fatty acid, sold under the trade names "SPAN 20, 40, 60 and 80" by the company "ICI"; - Sorbitan isostearate, sold under the trade name "SI 10 R NIKKOL" by the company "NIKKO"; sorbitan stearate bearing 4 ethylene oxide units, sold under the name "TWEEN 61" by the company "ICI"; polyethylene glycol stearate with 8 ethylene oxide units, sold under the name "MYR J 45" by the company "ICI"; the monostearate of polyethylene glycol of formula EMI6. 1 formula wherein n is 4, sold under the name "MYS 4" by the company "NIKKO"; polyethylene glycol stearate of molecular weight 400, chemical quality or quality produced by biotechnology, sold by the company "UNICHEMA"; - The diglyceryl stearate bearing 4 ethylene oxide units sold under the name "HOSTACERINE DGS" by the company "HOECHST"; tetraglycerol stearate, sold under the name TETRAGLYN 1S by the company "N I KKO"; the diglyceryl isostearate sold by the company "Solvay"; - The diglyceryl distearate, sold under the name "EMALEX DSG 2" by the company "NIHON"; - mono-, di- and tripalmitostearate sucrose, sold under the names "F50, F70, F110 and F160 CRODESTA" by the company "CRODA"; the mixture of sucrose mono- and dipalmitostearate, sold under the name "GRILLOTEN PSE 141 G" by the company "GRILLO"; the mixture of sucrose stearate and sucrose cocoate, sold under the name "Arlatone 2121" by the company "ICI"; the methylglucose distearate bearing 20 ethylene oxide units, sold under the name "Glucam E 20 DISTEARATE" by the company "AMERCHOL".

  Mixtures of these different products, which are already mixtures, or mixtures of these products with pure products, can of course be used.

  The cationic surfactants are selected from the list below and such that they give the dispersion a pH of between 5 and 8; the ratio by weight between the amount of nonionic amphiphilic lipids and that of cationic surfactants in the lipid phase being between 50/1 and 50/25 and the ratio by weight between the lipid phase and the aqueous dispersion phase being between 1 / 1000 and 300/1000.

  Oleosomes The oleosomes relating to the invention are described in the patent application EP 0 705 593. This is an oil-in-water emulsion formed of oily globules provided with a lamellar liquid crystal coating dispersed in an aqueous phase. , characterized in that each oily globule is unitarily coated with a monolamellar or oligolamellar layer (1 to 10 leaflets viewable by transmission electron microscopy after cryofracture) obtained from at least one lipophilic surfactant, at least one agent hydrophilic surfactant and at least one cationic surfactant imparting to the emulsion a pH of from 5 to 8, the coated oily globules having an average diameter of less than 500 nanometers.

  The lipophilic surfactant and the hydrophilic surfactant each comprise at least one saturated fatty chain having more than about 12 carbon atoms. More preferably still, this fatty chain contains from 16 to 22 carbon atoms.

  According to another preferred embodiment of the invention, the lipophilic surfactant has an HLB between about 2 and about 5. As is well known, the term HLB (Hydrophilic-Lipophilic Balance) means the balance between the dimension and the strength of the hydrophilic group and the size and lipophilic group strength of the surfactant.

  Examples of such lipophilic surfactants are sucrose distearate, diglyceryl distearate, tetraglyceryl tristearate, decaglyceryl decastearate, diglyceryl monostearate, hexaglyceryl tristearate, decaglyceryl pentastearate, sorbitan monostearate, and the like. sorbitan tristearate, diethylene glycol monostearate, glycerol ester of palmitic and stearic acid, polyoxyethylenated 2 EO monostearate (comprising 2 oxyethylene units), glyceryl mono and dibehenate, pentaerythritol tetrastearate.

  The hydrophilic surfactant preferably has an HLB of from about 8 to about 12.

  Examples of such hydrophilic surfactants are the following: polyoxyethylenesorbitan monostearate 40E, polyoxyethylenesorbitan tristearate 20OE, polyoxyethylene monostearate 80E, hexaglyceryl monostearate, polyoxyethylenated monostearate 10E, polyoxyethylenated distearate 12 EO and polyoxyethylenated methylglucose distearate 20 OE.

  The cationic surfactants may be chosen from the compounds mentioned below.

  Nanoemulsions The cationic particles may also be chosen from oil-water nanoemulsions comprising an oily phase dispersed in an aqueous phase, whose oil globules have a number average size of less than 100 nm, characterized in that they comprise at least an amphiphilic lipid comprising at least one nonionic amphiphilic lipid and a cationic amphiphilic lipid, the oily phase and the aminophilic lipids being present in a content such that the oily phase / amphiphilic lipid weight ratio ranges from 3 to 10. Nanoemulsions generally have a transparent to bluish appearance. Their

  transparency is measured by a transmittance coefficient at 600 nm ranging from 10 to 90% or by a turbidity. The turbidity of the compositions of the invention ranges from 60 to 400 NTU and preferably from 70 to 300 NTU, turbidity measured with the portable turbidimeter HACH - Model 2100 P at approximately 25 ° C.

  The oil globules of the nanoemulsions of the invention have a number average size, less than 100 nm and preferably ranging from 20 to 80 nm and more preferably from 40 to 60 nm. The decrease in the size of the globules makes it possible to promote the penetration of the active ingredients in the superficial layers of the skin (vehicle effect).

  The nanoemulsions according to the invention are preferably prepared at temperatures ranging from 4 to 45 ° C. and are thus compatible with heat-sensitive active substances.

  These dispersions are especially described in the following applications: EP 0728 460, EP 0 879 589, EP 1 010 413, EP 1 010 414, EP 1 010 416, EP 1 013 338, EP 1 016 453, EP 1 018 363, EP 1,025,898 and EP 1,120,102. In all these applications, it is specified that to improve the stability of the particles, an ionic surfactant will be added to the nonionic surfactant (or mixture).

  In the case of the present application, exclusively ionic surfactant will be used cationic surfactants or surfactants.

  The proportions indicated in the references above are to be preserved and as an example of a cationic surfactant, the list common to all the particles will be retained.

  The nonionic surfactants, preferably water-soluble or water-dispersible, comprise at least one hydrophobic block and at least one hydrophilic block.

  The nonionic amphiphilic lipids of the invention are preferably chosen from: 1 / silicone surfactants, 2 / liquid amphiphilic lipids at a temperature of less than or equal to 45 ° C., chosen from esters of at least one polyol of at least one fatty acid comprising at least one C8-C22 alkyl chain, saturated or unsaturated, linear or branched, and in particular unsaturated or branched, the polyol being chosen from the group formed by polyethylene glycol comprising from 1 to 60 oxide units; ethylene, sorbitan, glycerol may contain from 2 to 30 ethylene oxide units, polyglycerols having from 2 to 15 glycerol units, 3 / fatty acid esters of sugar and fatty alcohol ethers and sugar, 4 / solid surfactants at a temperature equal to 45 C selected from glycerol fatty esters, sorbitan fatty esters and oxyethylenated sorbitan fatty esters, ethoxylated fatty ethers and glycerol esters. ethoxylates, ethylene oxide (A) and propylene oxide (B) block copolymers, and mixtures of these surfactants.

  1 / The silicone surfactants that can be used according to the invention are silicone compounds comprising at least one oxyethylenated -OCH 2 CH 2 and / or oxypropylene-OCH 2 CH 2 CH 2 - chain, those described in documents US-A-5,364,633 and US-A-5,411,744.

  Preferably, the silicone surfactant used according to the present invention is a compound of formula (II): embedded image in which: R 1, R 2, R 3, independently of one another others represent a C 1 -C 6 alkyl radical or a radical - (CH 2) X (OCH 2 CH 2) y- (OCH 2 CH 2 CH 2) Z OR 4, at least one radical R 1, R 2 or R 3 not being an alkyl radical; R4 being a hydrogen, an alkyl radical or an acyl radical; A is an integer from 0 to 200; B is an integer from 0 to 50; provided that A and B are not equal to zero at the same time; x is an integer from 1 to 6; y is an integer from 1 to 30; z is an integer from 0 to 5.

  According to a preferred embodiment of the invention, in the compound of formula (X), the alkyl radical is a methyl radical, x is an integer ranging from 2 to 6 and y is an integer ranging from 4 to 30.

  Mention may be made, by way of example of silicone surfactants of formula (II), of the compounds of formula (III): (CH 3) 3 SiO - [(CH 3) 2 SiO] q - (CH 3 SO) g -Si (CH 3) 3 (CH2) 2- (OCH2CH2) y-OH wherein A is an integer from 20 to 105, B is an integer from 2 to 10 and y is an integer from 10 to 20.

  Mention may also be made, by way of example of silicone surfactants of formula (II), of the compounds of formula (IV): ## STR2 ## 3- (OCH2CH2) v-OH wherein A 'and y are integers ranging from 10 to 20.

  As silicone surfactants, those marketed by the company Dow Corning under the names DC 5329, DC 7439-146, DC 2-5695 and Q4-3667 may be used in particular. Compounds DC 5329, DC 7439-146, DC 2-5695 are compounds of formula (XI) where A is respectively 22, B is 2 and y is 12; A is 103, B is 10 and y is 12; A is 27, B is 3 and y is 12.

  Compound Q4-3667 is a compound of formula (IV) wherein A is and y is 13.

  2 / Amphiphilic lipids which are liquid at a temperature of less than or equal to 45 [deg.] C. may in particular be chosen from: polyethylene glycol isostearate of molar weight 400 (CTFA name: PEG-8 Isostearate), sold under the name Prisorine 3644 by the company Unichema; Diglyceryl isostearate, sold by Solvay; polyglycerol laurate comprising 2 units of glycerol (polyglyceryl-2 laurate), sold under the name Diglycerin-monolaurate by the company Solvay; Sorbitan oleate, sold under the name Span 80 by the company ICI; Sorbitan isostearate, sold under the name NIKKOL SI 10R by the company NIKKO; the α-butylglucoside cocoate or the α-butylglucoside caprate marketed by ULICE.

  3 / The fatty acid and sugar esters which can be used as nonionic amphiphilic lipids in the nanoemulsion according to the invention are preferably solid at a temperature of less than or equal to 45 ° C. and can be chosen in particular from the group comprising the esters or mixtures of C8-C22 fatty acid esters and sucrose, maltose, glucose or fructose, and esters or mixtures of C14-C22 fatty acid esters and methylglucose.

  The C8-C22 or C14-C22 fatty acids forming the fatty unit of the esters that can be used in the nanoemulsion of the invention comprise a linear saturated or unsaturated alkyl chain, having respectively 8 to 22 or 14 to 22 carbon atoms. . The fatty unit of the esters may in particular be chosen from stearates, behenates, arachidonates, palmitates, myristates, laurates, caprates and their mixtures. Stearates are preferably used.

  Examples of esters or mixtures of fatty acid esters and sucrose, maltose, glucose or fructose, sucrose monostearate, sucrose distearate, sucrose tristearate and the like. mixtures thereof, such as the products marketed by Croda under the name Crodesta F50, F70, F110, F160 respectively having a HLB (Hydrophilic Lipophilic Balance) of 5, 7, 11 and 16; and as an example of esters or mixtures of fatty acid esters and methylglucose, methyl glucose and polyglycerol-3 distearate, sold by Goldschmidt under the trade name Tego-care 450. also mention the monoesters of glucose or maltose such as o-hexadecanoyl-6D-glucoside methyl and o-hexadecanoyl-6-D-maltoside.

  The fatty alcohol and sugar ethers which can be used as nonionic amphiphilic lipids in the nanoemulsion according to the invention are solid at a temperature of less than or equal to 45 ° C. and can be chosen in particular from the group comprising ethers or ether mixtures. of C8-C22 fatty alcohol and glucose, maltose, sucrose or fructose and ethers or mixtures of C14-C22 fatty alcohol ethers and methylglucose. These are in particular alkylpolyglucosides.

  The C8-C22 or C14-C22 fatty alcohols forming the fatty unit of the ethers that can be used in the nanoemulsion of the invention comprise a saturated or unsaturated linear alkyl chain, having respectively 8 to 22 or 14 to 22 carbon atoms. . The fatty unit of the ethers may especially be chosen from decyl, cetyl, behenyl, arachidyl, stearyl, palmityl, myristyl, lauryl, capryl and hexadecanoyl units, and mixtures thereof such as cetearyl.

  Examples of fatty alcohol ethers and sugar include alkylpolyglucosides such as decylglucoside and laurylglucoside marketed for example by Henkel under the respective names of Plantaren 2000 and Plantaren 1200, cetostearylglucoside optionally in mixture with cetostearyl alcohol, sold for example under the name Montanov 68 by the company Seppic, under the name Tego-care CG90 by the company Goldschmidt and under the name Emulgade KE3302 by Henkel, and arachidylglucoside, for example in the form of a mixture of arachidic and behenic alcohols and arachidylglucoside, sold under the name Montanov 202 by the company Seppic.

  More particularly, nonionic amphiphilic lipids of this type are sucrose monostearate, sucrose distearate, sucrose tristearate and mixtures thereof, methyl glucose and polyglycerol-3 distearate and alkyl polyglucosides.

  4 / Fatty esters of glycerol, usable as nonionic amphiphilic lipids in the nanoemulsion according to the invention, solid at a temperature of less than or equal to 45 ° C., may be chosen in particular from the group comprising esters formed of at least one acid comprising a saturated linear alkyl chain having from 16 to 22 carbon atoms and from 1 to 10 glycerol units. One or more of these glycerol fatty esters may be used in the nanoemulsion of the invention.

  These esters may especially be chosen from stearates, behenates, arachidates and palmitates, and mixtures thereof. Stearates and palmitates are preferably used.

  As examples of surfactants that can be used in the nanoemulsion of the invention, mention may be made of decaglycerol monostearate, distearate, tristearate and pentastearate (10 units of glycerol) (CTFA names: Polyglyceryl-10 stearate, Polyglyceryl-10 distearate, Polyglyceryl 10-tristearate, Polyglyceryl-10 pentastearate) such as the products sold under the respective names Nikkol Decaglyn 1-S, 2-S, 3-S and 5-S by the company Nikko, and the diglyceryl monostearate (CTFA name: Polyglyceryl -2 stearate) such as the product sold by Nikko under the name Nikkol DGMS.

  The fatty esters of sorbitan, usable as nonionic amphiphilic lipids in the nanoemulsion according to the invention, which are solid at a temperature of less than or equal to 45 ° C., are chosen in particular from the group comprising the C 16 -C 22 fatty acid esters and from sorbitan and C16-C22 fatty acid esters and oxyethylenated sorbitan. They are formed of at least one fatty acid comprising at least one saturated linear alkyl chain, having respectively from 16 to 22 carbon atoms, and sorbitol or ethoxylated sorbitol. The oxyethylenated esters generally comprise from 1 to 100 ethylene oxide units and preferably from 2 to 40 ethylene oxide units (OE).

  These esters may especially be chosen from stearates, behenates, arachidates, palmitates, and mixtures thereof. Stearates and palmitates are preferably used.

  As an example of sorbitan fatty ester and of oxyethylenated sorbitan fatty ester, usable in the nanoemulsion of the invention, mention may be made of sorbitan monostearate (CTFA name: Sorbitan stearate) sold by the company ICI under the name Span 60 denomination, sorbitan monopalmitate (CTFA name: Sorbitan palmitate) sold by ICI under the name Span 40, sorbitan tristearate 20 OE (CTFA name: Polysorbate 65) sold by the company ICI under the name Tween 65.

  The ethoxylated fatty ethers which are solid at a temperature of less than or equal to 45 ° C., which can be used as nonionic amphiphilic lipids in the nanoemulsion according to the invention, are preferably ethers formed from 1 to 100 ethylene oxide units and at least a chain of fatty alcohol having from 16 to 22 carbon atoms. The fatty chain of the ethers may in particular be chosen from behenyl, arachidyl, stearyl and cetyl units, and mixtures thereof such as cetearyl. By way of example of ethoxylated fatty ethers, mention may be made of behenic alcohol ethers comprising 5, 10, 20 and 30 ethylene oxide units (CTFA names: Beheneth-5, Beheneth-10, Beheneth-20 , Beheneth-30), such as the products marketed under the names Nikkol BB5, BB10, BB20, BB30 by the company Nikko, and the stearyl alcohol ether comprising 2 ethylene oxide units (CTFA name: Steareth- 2), such as the product sold under the name Brij 72 by the company ICI.

  The solid ethoxylated fatty esters at a temperature of less than or equal to 45 ° C., which can be used as nonionic amphiphilic lipids in the nanoemulsion according to the invention, are esters formed from 1 to 100 ethylene oxide units and from at least one chain. of fatty acid having from 16 to 22 carbon atoms. The fatty chain of the esters may especially be chosen from stearate, behenate, arachidate and palmitate units, and mixtures thereof. By way of example of ethoxylated fatty esters, mention may be made of the stearic acid ester comprising 40 ethylene oxide units, such as the product sold under the name Myrj 52 (CTFA name: PEGstearate) by the company ICI as well as the behenic acid ester comprising 8 ethylene oxide units (CTFA name: PEG-8 behenate), such as the product sold under the name Compritol HD5 ATO by the company Gattefosse.

  5 / The block copolymers of ethylene oxide and of propylene oxide, usable as nonionic amphiphilic lipids in the nanoemulsion according to the invention may be chosen in particular from block copolymers of formula (V): HO (C2H40) x (C3H60) y (C2H40) zH (V) wherein x, y and z are integers such that x + z is from 2 to 100 and y is from 14 to 60, and mixtures thereof, and more particularly from copolymers blocks of formula (V) having an HLB ranging from 2 to 16.

  These block copolymers may especially be chosen from poloxamers and in particular from Poloxamer 231 such as the product sold by the company ICI under the name Pluronic L81 of formula (V) with x = z = 6, y = 39 (HLB 2); Poloxamer 282 such as the product sold by ICI under the name Pluronic L92 of formula (V) with x = z = 10, y = 47 (HLB 6); and Poloxamer 124, such as the product sold by ICI under the name Pluronic L44 of formula (V) with x = z = 11, y = 21 (HLB 16).

  As nonionic amphiphilic lipids, mention may also be made of the mixtures of nonionic surfactants described in the document EP-A-705593 incorporated herein by reference.

  Among the nonionic amphiphilic lipids, it is possible to use in particular: PEG 400 or PEG-8 isostearate Isostearate (comprising 8 moles of ethylene oxide), diglyceryl isostearate, polyglycerol monolaurate comprising 2 units of glycerol and polyglycerol stearates containing 10 units of glycerol, sorbitan oleate, sorbitan isostearate, and mixtures thereof.

  The nonionic amphiphilic lipids may be present in the nanoemulsion according to the invention in a content ranging from 0.2% to 12% by weight, relative to the total weight of the composition, and preferably ranging from 0.2% to 8% by weight. % by weight, and preferably ranging from 0.2% to 6% by weight.

  The cationic amphiphilic lipids are chosen from the list given below.

  They are present in the nanoemulsions of the invention, preferably in concentrations ranging from 0.01 to 6% by weight relative to the total weight of the nanoemulsion and more particularly from 0.2 to 4% by weight.

  Oils: The oily phase of the nanoemulsion according to the invention comprises at least one oil. The oils which can be used in the nanoemulsions of the invention are preferably chosen from the group formed by: oils of animal or vegetable origin, formed by esters of fatty acids and of polyols, in particular liquid triglycerides, for example, sunflower, corn, soybean, avocado, jojoba, squash, grape seed, sesame, hazelnut, fish oil, tricaprocaprylate glycerol, or vegetable or animal oils of the formula R9000R10 in which R9 represents the residue of a higher fatty acid having from 7 to 29 carbon atoms and R10 represents a linear or branched hydrocarbon chain containing from 3 to 30 carbon atoms, in particular alkyl or alkenyl, for example, Purcellin oil or liquid jojoba wax; natural or synthetic essential oils such as, for example, eucalyptus, lavandin, lavender, vetiver, litsea cubeba, lemon, sandalwood, rosemary, chamomile, savory, walnut nutmeg, cinnamon, hyssop, caraway, orange, geraniol, cade and bergamot; synthetic oils such as parleam oil, polyolefins and liquid carboxylic acid esters; mineral oils such as hexadecane, isohexadecane and paraffin oil; halogenated oils, in particular fluorocarbons such as fluoroamines, for example perfluorotributylamine, fluorinated hydrocarbons, for example perfluorodecahydronaphthalene, fluoroesters and fluoroethers; volatile or non-volatile silicone oils.

  The polyolefins that may be used as synthesis oils are in particular poly-α-olefins and more particularly those of polybutene type, hydrogenated or otherwise, and preferably polyisobutene, hydrogenated or otherwise.

  The liquid carboxylic acid esters which can be used as synthesis oils may be esters of mono-, di-, tri- or tetracarboxylic acids. The total number of carbon of the esters is generally greater than or equal to 10 and preferably less than 100 and more particularly less than 80. It is especially the monoesters of saturated or unsaturated aliphatic acids, linear or branched C 1 -C 26 and of saturated or unsaturated aliphatic alcohols, linear or branched C1-C26, the total number of carbon esters being generally greater than or equal to 10. It is also possible to use the esters of di- or tri-carboxylic acids C4-C22 and d C1-C22 alcohols and esters of mono-, di- or tri-carboxylic acids and di-, tri-, teta- or penta-hydroxylated C2-C26 alcohols.

  Among the esters mentioned above, it is preferred to use alkyl palmitates such as ethyl palmitate, isopropyl palmitate, ethyl-2-hexyl palmitate and 2-octyldecyl palmitate; alkyl myristates such as isopropyl myristate, butyl myristate, cetyl myristate, 2-octyldodecyl myristate; alkyl stearates such as hexyl stearate, butyl stearate, isobutyl stearate; alkyl malates such as dioctyl malate; alkyl laurates such as hexyl laurate and hexyl decyl laurate; Isononyl isononanate; cetyl octanoate.

  Advantageously, the nanoemulsion according to the invention contains at least one oil with a molecular weight greater than or equal to 400, in particular ranging from 400 to 10,000, better still from 400 to 5,000, or even from 400 to 2,500. Molecular weight oils greater than or equal to 400 may be chosen from oils of animal or vegetable origin, mineral oils, synthetic oils and silicone oils, and mixtures thereof. Examples of such oils are isoketyl palmitate, isoketyl stearate, avocado oil and jojoba oil.

  The nanoemulsions in accordance with the invention comprise a quantity of oily phase (oil and other fatty substances, excluding the amphiphilic lipid) preferably ranging from 2 to 40% by weight relative to the total weight of the nanoemulsion and more particularly from 4 to 30. % by weight and preferably from 4 to 20% by weight.

  The oily phase and the amphiphilic lipids (nonionic and ionic amphiphiles) are preferably present in the nanoemulsion according to the invention in a weight ratio of the amount of oily phase to the amount of amphiphilic lipid ranging from 3 to 10 and preferably ranging from 3 to 6. The term "amount of oily phase" here means the total amount of the constituents of this oily phase without including the amount of amphiphilic lipid.

  The nanoemulsions according to the present invention may contain, in addition to the urea derivatives of formula (I) described above, solvents, in particular to improve, if necessary, the transparency of the composition.

  These solvents are preferably chosen from the group formed by: lower C 1 -C 8 alcohols such as ethanol; glycols such as glycerin, propylene glycol, 1,3-butylene glycol, dipropylene glycol, polyethylene glycols containing from 4 to 16 ethylene oxide units and preferably from 8 to 12; sugars such as glucose, fructose, maltose, lactose, sucrose.

  These solvents can be used in a mixture. When they are present in the nanoemulsion of the invention, they can be used at concentrations ranging preferably from 0.01 to 30% by weight relative to the total weight of the nanoemulsion, and better still from 5 to 20% by weight. relative to the total weight of the nanoemulsion. The amount of alcohol (s) and / or sugar (s) is preferably from 5 to 20% by weight relative to the total weight of the nanoemulsion and the amount of glycol (s) is preferably from 5 to 15% by weight relative to the total weight of the nanoemulsion.

  Preparation process: The process for preparing a nanoemulsion as defined above consists in mixing the aqueous phase containing the urea derivative and the oily phase, with vigorous stirring, at a temperature ranging from 10 ° C. to 80 ° C., to perform a high pressure homogenization step at a pressure greater than 5.107 Pa and to optionally add the polymer used. According to a preferred embodiment of the invention, a high pressure homogenization step is then further carried out at a pressure greater than 5.107 Pa. The high pressure homogenization is preferably carried out at a pressure ranging from 6.107 to 18.107 Pa. Preferably, the shear is from 2.106 to 5.108, and more preferably from 1 × 10 8 to 3.108 (s = second-1) .This method makes it possible to produce nanoemulsions which are compatible with heat-sensitive active compounds and which may contain oils and especially perfumes which contain fatty substances, without denaturing them.

  Nanocapsules The nanocapsules relating to the invention are those described in patent applications EP 0 447 318, EP 0 557 489, EP 0 780 115, EP 1 025 901, EP 1 029 587, EP 1 034 839 and EP 1 414 390. , FR 2 830 776, EP 1 342 471, FR 2 848 879 and FR 04/50057.

  Nanocapsules are core-shell particles having an oily core and a polymeric shell. The various applications cited above relate to different families of polymers and different processes for obtaining them. The size of the capsules is always less than 1 μm and it is possible to have sizes smaller than 80 nm. These particles may be coated with a lamellar liquid crystal phase, most often consisting of a lecithin or a dimethicone copolyol. The coating must consist of an amphiphilic lipid capable of spontaneously forming a lamellar liquid crystal phase in contact with water. It is to this amphiphilic lipid capable of forming a lamellar phase that the cationic surfactant will be added which will give the particles (the nanocapsule) a positive zeta potential. The weight ratio between the amphiphilic lipid forming the lamellar phase and the cationic surfactant will be between 99/1 and 75/25.

  The cationic surfactants that can be used are those listed below.

  Organic particles The organic particles of the invention are nanospheres, solid, without internal cavity formed by different processes (dispersion in water, nanoprecipitation, microemulsion ...) and composed of at least one polymer or at least one copolymer or a mixture thereof. The particle is cationic, with the zeta potential defined above, either because the polymer (s) or copolymer (s) are cationic or nonionic and a cationic surfactant as described below is used. With respect to the polymer, the level of cationic surfactant will be between 0 and 25%.

  Inorganic particles The cationic inorganic particles of the invention may be, for example, based on silica, TiO2, ZnO, alumina, etc. By way of example, mention may be made of alumina particles in colloidal dispersion. in the water like the Nanomer 2 of Nalco. Clariant and Grace also offer particles of this type.

  Cationic surfactants that can be used for the preparation of the cationic particles of the invention The cationic surfactants that can be used according to the invention are listed below, this list is not limiting.

  The cationic amphiphilic lipids are preferably chosen from the group consisting of quaternary ammonium salts, fatty amines and their salts.

  The quaternary ammonium salts are, for example: those having the following general formula (IV): + R1.

NOT

r + '' -.

R 2 4

  in which the radicals R1 to R4, which may be identical or different, represent a linear or branched aliphatic radical containing from 1 to 30 carbon atoms, or an aromatic radical such as aryl or alkylaryl. The aliphatic radicals can comprise heteroatoms such as in particular oxygen, nitrogen, sulfur, halogens. The aliphatic radicals are, for example, chosen from alkyl, alkoxy, polyoxyalkylene (C 2 -C 6), alkylamide, (C 12 -C 22) alkylamido (C 2 -C 6) alkyl, (C 12 -C 22) alkyl acetate and hydroxyalkyl radicals containing from about 1 to 30 carbon atoms; X is a

X

  anion chosen from the group of the halides, phosphates, acetates, lactates, (C 2 -C 6) alkyl sulphates, alkyl or alkylarylsulphonates, and the quaternary ammonium salts of imidazolinium, for example that of the following formula (V) in which R5 represents an alkenyl or alkyl radical containing from 8 to 30 carbon atoms, for example derived from tallow fatty acids, R6 represents a hydrogen atom, a C1-C4 alkyl radical or an alkenyl or alkyl radical containing 8 to 30 carbon atoms, R7 represents a C1-C4 alkyl radical, R8 represents a hydrogen atom, a C1-C4 alkyl radical, X is an anion chosen from the group of halides, phosphates, acetates, lactates, alkylsulfates, alkyl-or-alkylarylsulfonates. Preferably, R 5 and R 6 denote a mixture of alkenyl or alkyl radicals containing from 12 to 21 carbon atoms, for example derived from tallow fatty acids, R 7 denotes methyl and R 8 denotes hydrogen. Such a product is, for example, sold under the name REWOQUAT W 75 by the company REWO, the quaternary diammonium salts of formula (VI): ## STR2 ## in which R 9 denotes an aliphatic radical containing approximately from 16 to 30 carbon atoms, R10, R11, R12, R13 and R14, which are identical or different, are chosen from hydrogen or an alkyl radical containing from 1 to 4 carbon atoms, and X is an anion chosen from the group of halides, acetates, phosphates, nitrates and Such diammonium quaternary salts include, in particular, propane diammonium dichloride: quaternary ammonium salts containing at least one ester function The quaternary ammonium salts containing at least one ester function which can be used according to the invention are for example those of the following formula (VII): ## STR5 ## in which: R 15 is chosen from C 1 -C 6 alkyl radicals and C 1 -C 6 hydroxyalkyl or dihydroxyalkyl radicals, R 16 is among: - the radical - linear or branched, saturated or unsaturated C1-C22 hydrocarbon radicals R20 - hydrogen atom, - R18 is chosen from: - the radical - linear C1-C6 hydrocarbon radicals R22 or branched, saturated or unsaturated, - the hydrogen atom, - R17, R19 and R21, which are identical or different, are chosen from linear or branched, saturated or unsaturated C7-C21 hydrocarbon radicals; n, p and r, which may be identical or different, are integers ranging from 2 to 6; y is an integer from 1 to 10; x and z, identical or different, are integers ranging from 0 to 10; - X- is a simple or complex anion, organic or inorganic; with the proviso that the sum x + y + z is from 1 to 15, that when x is 0 then R16 is R20 and that when z is 0 then R18 is R22.

  The alkyl radicals R15 may be linear or branched and more particularly linear.

  Preferably R15 denotes a methyl, ethyl, hydroxyethyl or dihydroxypropyl radical and more particularly a methyl or ethyl radical. Advantageously, the sum x + y + z is from 1 to 10.

  When R 16 is a hydrocarbon radical R 20, it can be long and have 12 to 22 carbon atoms or short and have 1 to 3 carbon atoms.

  When R 18 is a hydrocarbon radical R 22, it preferably has 1 to 3 carbon atoms. Advantageously, R17, R19 and R21, which are identical or different, are chosen from linear or branched, saturated or unsaturated C11-C21 hydrocarbon radicals, and more particularly from linear or branched, saturated or saturated C11-C21 alkyl and alkenyl radicals. or unsaturated.

  Preferably, x and z, identical or different, are 0 or 1.

Advantageously, y is 1.

  Preferably, n, p and r, identical or different, are 2 or 3 and even more particularly are equal to 2.

  The anion is preferably a halide (chloride, bromide or iodide) or an alkyl sulphate more particularly methyl sulphate. However, it is possible to use methanesulphonate, phosphate, nitrate, tosylate, an anion derived from an organic acid such as acetate or lactate or any other anion compatible with ammonium with an ester function.

  The X- anion is even more particularly chloride or methylsulfate.

  More particularly, the ammonium salts of formula (VII) in which: R15 denotes a methyl or ethyl radical, - x and y are equal to 1; z is 0 or 1; n, p and r are 2; - R16 is chosen from: - the radical 0 Il - methyl, ethyl or hydrocarbon radicals C14-C22 - the hydrogen atom; - R18 is chosen from: - the radical the hydrogen atom; R17, R19 and R21, which are identical or different, are chosen from linear or branched, saturated or unsaturated C13-C17 hydrocarbon radicals and preferably from linear or branched, saturated or unsaturated C13-C17 alkyl and alkenyl radicals.

  Advantageously, the hydrocarbon radicals are linear.

  There may be mentioned for example the compounds of formula (VII) such as the salts (especially chloride or methylsulfate) of diacyloxyethyl dimethyl ammonium, diacyloxyethyl hydroxyethyl methyl ammonium, monoacyloxyethyl dihydroxyethyl methyl ammonium, triacyloxyethyl methyl ammonium, monoacyloxyethyl hydroxyethyl dimethyl ammonium and their mixtures. The acyl radicals preferably have from 14 to 18 carbon atoms and come more particularly from a vegetable oil such as palm oil or sunflower oil.

  When the compound contains more than one acyl radical, the latter may be identical or different.

  These products are obtained for example by direct esterification of triethanolamine, triisopropanolamine, alkyldiethanolamine or alkyldiisopropanolamine optionally oxyalkylenated on fatty acids or mixtures of fatty acids of plant or animal origin or by transesterification of their esters methyl. This esterification is followed by a quaternization using an alkylating agent such as an alkyl halide (preferably methyl or ethyl), a dialkyl sulphate (methyl or preferably ethyl), methyl methanesulphonate, methyl paratoluenesulfonate, glycol or glycerol chlorohydrin.

  Such compounds are for example sold under the names DEHYQUART by Henkel, STEPANQUAT by STEPAN, NOXAMIUM by CECA, REWOQUAT WE 18 by REWOWITCO.

  The composition according to the invention preferably contains a mixture of quaternary ammonium mono, di and triester salts with a majority by weight of diester salts.

  As a mixture of ammonium salts, it is possible to use, for example, the mixture containing 15 to 30% by weight of acyloxyethyl dihydroxyethyl methyl ammonium methyl sulphate, 45 to 60% of diacyloxyethyl hydroxyethyl methyl ammonium methyl sulphate and 15 to 30% of methyl sulphate. of triacyloxyethyl methyl ammonium, the acyl radicals having from 14 to 18 carbon atoms and coming from partially hydrogenated palm oil. The ammonium salts containing at least one ester function described in US Pat.

  Of the quaternary ammonium salts of formula (IV), tetraalkylammonium chlorides, for example dialkyldimethylammonium or alkyltrimethylammonium chlorides, in which the alkyl radical contains approximately from 12 to 22 carbon atoms, are preferred. , in particular behenyltrimethylammonium, distearyldimethylammonium, cetyltrimethylammonium, benzyl dimethylstearylammonium chlorides or, on the other hand, stearamidopropyldimethyl (myristyl acetate) ammonium chloride sold under the name CERAPHYL 70 by VAN DYK.

  According to the invention, behenyltrimethylammonium chloride or bromide and CTAB (cetyltrimethylammonium bromine) are the most particularly preferred quaternary ammonium salts.

  The fatty amines of the invention correspond to the general formula: RICONH (CH2) mNR2 R3-R is a saturated or unsaturated hydrocarbon chain, branched or unbranched, having between 8 and 30 carbon atoms and preferably between 10 and 24.

  - R2 and R3 are independently selected from saturated or unsaturated hydrocarbons, branched or unbranched, having between 1 and 10 carbon atoms and preferably between 1 and 4.

  - R2 and R3 may also, always independently of each other correspond to a hydrogen atom H. - M is between 1 and 10 and preferably between 1 and 5.

  By way of nonlimiting example, include: stearylamine, the aminoethylethanolamide Stearate, Stearyl diethanolamide, diethylenetriamine stearate, stearamidopropyldimethylamine, the stearamidopropyldiethylamine, stearamidoethyldiethylamine the the stearamidoethyldimethylamine, palmitamidopropyldimethylamine the the palmitamidopropyldiethylamine, palmitamidoethyldiethylamine the the palmitamidoethyldimethylamine, behenamidopropyldimethylamine the , behenamidopropyldiethylmine, behenamidoethyldiethylamine, behenamidoethyldimethylamine, arachidamidopropyldimethylamine, arachidamidopropyldiethylamine, arachidamidoethyldiethylamine, arachidamidoethyldimethylamine.

  Commercially available fatty amine include Croda Incromine BB, Nikkol Amidoamine MSP, Inolex Lexamine.

  Other fatty amines that may be mentioned include, for example, stearylamine, stearate aminoethylethanolamide, stearyl diethanolamide, stearate diethylenetriamine which, among others, Sabo works with the Sabomina series.

  Also noteworthy are fatty amine acetates such as the Acetamine series of Kao Corp. These fatty amines can also be ethoxylated, such as Berol 380, 390, 453 and 455, Ethomeen from Akzo Nobel or Marlazin L10, OL2, OL20, T15 / 2 and T50 from Condea Chemie.

  The particles of the present invention can be introduced into all galenic carriers for cosmetic, dermatological and ophthalmic purposes. By way of example, mention may be made of lotions, serums, gels and all types of emulsions.

  The compositions according to the invention may have all the galenical forms normally used for topical application, for example in the form of solutions, gels, dispersions of the lotion or serum type, emulsions of liquid or semi-liquid consistency of the milk type. obtained by dispersion of a fatty phase in an aqueous phase (O / W) or conversely (W / O), or suspensions or emulsions of soft, semi-solid or solid consistency of the cream or gel type, or even of microemulsions , microcapsules, microparticles or vesicular dispersions of ionic and / or nonionic type. These compositions are prepared according to the usual methods.

  The compositions according to the invention may furthermore comprise any additive usually used in the cosmetic or pharmaceutical field.

  Of course those skilled in the art will take care to choose this or these optional additional compounds, and / or their amount, such that the advantageous properties of the composition according to the invention are not, or not substantially impaired.

  In particular, it will be ensured that this introduction does not affect the stability of the cationic particles associated with siRNA.

  The composition according to the invention may in particular contain cosmetic or pharmaceutical active ingredients. Preferably, it will be depigmenting agents, sunscreens.

  Depigmenting agents that may be incorporated into the composition include, for example, the following compounds: kojic acid; ellagic acid; Arbutin and its derivatives such as those described in EP-895,779 and EP-524,109; hydroquinone; aminophenol derivatives such as those described in applications WO 99/10318 and WO 99/32077, and in particular N-cholesteryloxycarbonylparaaminophenol and N-ethyloxycarbonyl-para-aminophenol; iminophenol derivatives, in particular those described in application WO 99/22707; L-2-oxothiazolidine-4-carboxylic acid or procysteine, as well as its salts and esters; ascorbic acid and its derivatives, especially ascorbyl glucoside; and plant extracts, in particular licorice, mulberry and skullcap, without this list being limiting.

  The ultraviolet radiation filtering agents may be chosen from organic UV screening agents or mineral UV radiation filtering agents.

  The organic UV filters in accordance with the invention may be water-soluble, fat-soluble or insoluble in the usual cosmetic solvents. They are chosen in particular from anthranilates; cinnamic derivatives; dibenzoylmethane derivatives; salicylic derivatives, camphor derivatives; triazine derivatives such as those described in patent applications US 4,367,390, EP 863,145, EP 517,104, EP 570,838, EP 796,851, EP 775,698, EP 878,469 and EP 933,376; benzophenone derivatives, in particular those described in applications EP 1 046 391 and DE10012408; derivatives of f3, (3'-diphenylacrylate, benzotriazole derivatives, benzimidazole derivatives, imadazolines, bis-benzoazolyl derivatives as described in patents EP 669 323 and US 2,463,264; aminobenzoic acid (PABA), the bis (hydroxyphenylbenzotriazole) methylene derivatives as described in US 5,237,071, US 5,166,355, GB 2303549, DE 19726184 and EP 893 119, the filter and silicone filter polymers such as those described in particular in US Pat. application W093 / 04665, dimeres derived from alkylstyrene such as those described in the patent application DE19855649, 4,4-diarylbutadiene derivatives such as those described in patent applications EP 0 967 200 and DE19755649.

  The inorganic filtering agents are generally pigments or even nanopigments (average size of the primary particles: generally between 5 nm and 100 nm, preferably between 10 nm and 50 nm) of coated or uncoated metal oxides, for example nano-sized nanopigments. titanium oxide (amorphous or crystallized in rutile and / or anatase form), iron, zinc, zirconium or cerium which are all UV photoprotective agents well known per se. Conventional coating agents are, moreover, alumina and / or aluminum stearate. Such nanopigments of metal oxides, coated or uncoated, are in particular described in patent applications EP 0 518 772 and EP0518773.

  The radiation screening agents according to the invention are generally present in the compositions according to the invention in proportions ranging from 0.1 to 20% by weight relative to the total weight of the composition, and preferably ranging from 0.2 to 15% by weight relative to the total weight of the composition.

  According to another of its objects, the present invention also relates to the use of at least one double-stranded RNA oligonucleotide of sequence chosen from SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 and 48 to inhibit expression of tyrosinase.

  In particular, the oligonucleotides according to the invention are useful as whitening agents and / or anti-browning agents for the skin.

  The double-stranded RNA oligonucleotides according to the invention can also be used for the preparation of a topical composition, cosmetic or dermatological, intended to reduce the synthesis of melanin, in particular, with a view to treating hyperpigmentation, pigmentary tasks and dyschromias. or for the bleaching of hair follicles, regional hyperpigmentations by melanocytic hyperactivity such as idiopathic melasmas, occurring during pregnancy ("pregnancy mask" or chloasma) or estrogen-progestative contraception, hyperpigmentations localized by hyperactivity and proliferation benign melanocyte, such as senile pigmentary spots known as actinic lentigo, accidental hyperpigmentations, possibly due to photosensitization or post-injury healing, as well as certain leukoderma, such as vitiligo. For the latter (the scarring may result in a scar giving the skin a whiter appearance), failing to repigment the damaged skin, it completes depigmenting the areas of residual normal skin to give the whole skin a homogeneous white color.

  Another subject of the present invention relates to a cosmetic process for whitening and / or lightening the complexion and / or uniforming the color of a burnished skin comprising the topical application of at least one sequence double-stranded RNA oligonucleotide chosen from the SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 and 48.

  Preferably, the oligonucleotide is formulated in a topical composition at a concentration of less than or equal to 1 nM.

  In a still preferred manner, the oligonucleotide is associated with a cationic particle having a size of less than or equal to 1 μm, a zeta potential of between 10 and 80 mV chosen from surfactant micelles, block polymer micelles, liposomes of nonionic and cationic surfactants, niosomes, oleosomes, nanoemulsion particles, nanocapsules, organic particles or inorganic particles, as described above.

  For purposes of cosmetic, dermatological, ophthalmic or pharmaceutical care, it is sought to avoid invasive methods such as subcutaneous or ocular injections. The cationic particles associated with the siRNA according to the present invention can be applied topically to the skin, the mucous membranes or in the eye, in suspension or incorporated in an acceptable cosmocentric support, such as lotions, serum, emulsified gels or not, oil emulsions. / water, water / oil, multiples, microemulsions ...

  To promote the penetration of the particles associated with siRNA, we can change the permeability of the skin, so it is possible to control the permeability of the skin by measuring the PIE (insensible loss of water) with a Tewametre TM210 or a Dermalab Cortex technology.

  By way of example, the following methods may be used: stripping (corneodisc, varnish), chemical peel or mechanical dermabrasion before application of the composition according to the invention; pre-treatment with a mixture of one or more solvents having a delipidating effect; cleaning the skin with a detergent foaming product; -pré and / or post occlusive treatment is for example by covering the surface of the skin to be treated with a tight synthetic membrane (Blenderm for example) or by the application of a layer of Vaseline. This has the effect of blocking the skin's natural PIE and causing over-hydration of the epidermal lipids which thus become more permeable.

  After the application of the composition of the present invention, one can also use methods known to those skilled in the art favoring the penetration of molecules within the skin such as, for example, iontophoresis or electroporation.

  In the case of an in vitro skin model, the following treatments may be applied: - before deposition of the composition according to the invention, it will be possible to reduce the level of Ca 2+ either by changing the medium (0.100.75 mM instead of 1.5 mM) or by adding a chelant. This has the effect of reducing intermembrane adhesion, thus increasing the permeability of the skin (Effects of extra- and intracellular calcium concentration on DNA replication, lateral growth, and differentiation of human epidermal cells in culture.) Virchows Arch B Cell Pathol lncl Mol Pathol 1990, 59 (1): 17-25); using iontophoresis, electroporation and any known methods for modifying the permeability of the model.

  EXAMPLE 1 - Cationic Micelle Formulations - Example 1: Octyl 3 Glucoside / cetyl triammonium bromide at a molar ratio of 5/1 El: 100 mM in distilled water E2: 50 mM in distilled water E3: 25 mM in distilled water E4: 12.5 mM in distilled water The 2 surfactants (nonionic + cationic) are solubilized in distilled water. A suspension of siRNA from SEQ. ID. 1 (20 NM) is added between 1: 1 and 1: 9 volume (siRNA: micelles) thus reducing the micellar concentration in proportion.

  Example 2 Micelles of decyl R glucoside / behenyl triammonium chloride at a molar ratio of 5/1 in distilled water. The same concentrations as in the previous example are realized.

  These 2 suspensions can be applied to the skin for the treatment of pigmentary spots and dyschromias or for whitening of hair follicles.

  Cationic liposomes - Example 1: fluid vesicles Isostearate PEG 400 5.5% Behenyl triammonium chloride 0.5% Distilled water qs 100 Example 2: Rigid vesicles 2.75% Sorbitan palmitate Cholesterol 2.75% Behenyl triammonium chloride 0.5% Distilled water qs 100 These examples of suspensions of Liposomes are made by dialysis. The constitutive lipids of the vesicles are solubilized in an aqueous solution of octyl glucoside. This solution is then dialyzed against water for 72 hours.

  The suspension of siRNA SEQ. ID. # 4 is then added, reducing the vesicle concentration around 3% lipid. This suspension of nonionic liposomes can be applied to the skin for the treatment of pigmentary stains and stains or for whitening of hair follicles.

  Cationic oleosomes - Sucrose oil phase Mono-di stearate sold by Stéarainerie Dubois 0.45% Sorbitan stearate 40E (Tween 61 Uniquema) 0.30% Behenyl triammonium chloride 0.21% Vitamin E acetate 0.5% Jojoba oil 0.5% Stearyl heptanoate 1% Oil volatile silicone 1% Vit F glyceride 0.5% Preservative 0.02% BHT 0.01% 30 Aqueous phase Distilled water qs 100 0.1% preservative This dispersion is produced by high pressure homogenization in order to have a particle size of about 170 nm. Then added is a suspension of siRNA SEQ. ID. 16 (20 NM) at ratii ranging from 1: 1 to 1:20 (siRNA / oleosomes). The siRNA will then complex on the surface of the particles.

  Cationic nanocapsules - Organic phase 1% Polycaprolactone (MW: 50000) Vit E 1% Dimethicone copolyol DC2 5695 (Dow Corning) 0.5% Behenyl triammonium chloride 0.21% Acetone 200 ml - Aqueous phase 0.5% Pluronic F68 Distilled water 200ml The organic phase is introduced with stirring in the aqueous phase. Acetone and 100 ml of aqueous phase are then evaporated to obtain the suspension of nanocapsules whose size is 220 nm. Then added is a suspension of siRNA SEQ. ID. No. 37 (20 μM) at ratii ranging from 1: 1 to 1:20 (siRNA / nanocapsules). The siRNA will then complex on the surface of the particles.

  Organic Nanoparticles - Organic Phase Polyethylene Adipate (Scientific Polymer products) 2% Dimethicone copolyol DC2 5695 (Dow Corning) 0.5% Behenyl triammonium chloride 0.21% Acetone 200 ml - aqueous phase Pluronic F68 0.5% Distilled water 200 ml The organic phase is stirred into the aqueous phase. Acetone and 100 ml of aqueous phase are then evaporated to obtain the suspension of nanoparticles whose size is 180 nm. Then added is a suspension of siRNA SEQ. ID. No. 40 (20 μM) at ratii ranging from 1: 1 to 1:20 (siRNA / nanoparticles). The siRNA will then complex on the surface of the particles.

  Cationic Nanoemulsions - Oil Phase PEG 400 Isostearate sold by Uniqema 35 Behenyl triammonium chloride Avocado oil Jojaba oil Cyclopentamethylsiloxane 2% - Water phase 30% Distilled water Dipropylene glycol 10% - Dilution phase gsp100% Distilled water Preservative 0.1% An emulsion is carried out by dispersing the oily phase on the aqueous phase with very vigorous stirring. The suspension obtained is then homogenized several times using a very high pressure homogenizer, at a pressure of about 1200 b. The particle size is of the order of 50 nm and the suspension is transparent. The dilution phase is then added.

  As in the previous cases, a suspension of siRNA SEQ is then added. ID. No. 42 (20 μM) at ratii ranging from 1: 1 to 1:20 (siRNA / nanoemulsion). The siRNA will then complex on the surface of the particles.

  EXAMPLE 2 Measurement of SiRNA Activity According to the Invention The efficacy of the 47 siRNAs of sequence SEQ ID No. 1 to 47 according to the invention was evaluated in the pSCREEN-iTTM system test developed by Invitrogen developed to evaluate the efficiency on tyrosinase inhibition (GenBank accession number M27160).

  Experimental Test Protocol: Seeding of 2,104 A549 cells / well in a 48-well plate and grown overnight.

  Co-transfection in triplicate for 5 h using 2 μg / ml of Lipofectamine 2000 (Invitrogen) complexed with Tyrosinase pScreen-iT ™ vector coupled to LacZ gene 10 ng / well Luciferase reporter plasmid 20 ng / well É 1 nM StealthTM RNA (sequences Data in Table I) The cells are lysed 24h after transfection and the Galactosidase (8-Gal) and Luciferase (Luc) activities are measured. The ratio [3Gal / Luc determines the percent inhibition of tyrosinase.

  The results obtained are shown in Table II.

  Table II: Assay of the Efficacy of 47 Double-Stranded RNA Oligonucleotides on Tyrosinase mRNA Degradation (GenBank accession number M27160).

  Oligonucleotide Oligonucleotide (SEQ ID NO)% Inhibition (SEQ ID NO)% Inhibition 1 97.50% 22 77.91% 2 98.57% 3 86.95% 4 97.95% 94.17% 6 77, 68% 7 95.86% 97.50% 87.14% 91.54% 88.44% 89.50% 98.15% 14 97.87% 36.02% 97.78% 17 88.54 % 18 81.11% 83.33% 92.64% 43 99.10% 83.54% 89.05% 23 97, 29% 24 90.50% 95.17% 90.60% 87.11% 91 , 58% 29 96.00% 90.39% 31 10.38% 32 97.93% 84.79% 64.43% 97.71% 92.12% 97.07% 38 9.55% 39 90, 22% 97.44% 88, 11% 98.71% 46 98.64% 94.14% 11 12 13 26 27 28 33 34 In a second step, a dose effect was performed on the 20 most effective sequences. with an efficiency greater than 91% at 1 nM. The results are shown in Table III.

  Table III: Assay of the Efficacy of the Selected Sequences by Dose Effect of 0.0016 nM to 1 nM.

  Oligonucleotide 1 nM 0.25 nM 0.063 nM 0.016 nM (SEQ ID NO: 1) 97.37% 94, 34% 89.47% 72.07% 2 97.98% 95.64% 89.34% 66.68% 4 97.93% 95.22% 90.24% 57, 92% 91.33% 79.88% 61.47% 18.40% 7 96.86% 88.97% 76.45% 38.34% 8 97.84% 90, 76% 81.12% 38.57% 13 98.13% 95.73% 91.11% 62.56% 14 97.59% 87.22% 68.77% 33.87 % 16 97.95% 94.92% 83.40% 56.25% 23 96.05% 87.42% 62.32% 29.97% 93, 45% 90.50% 78.36% 46.80 % 29 96.42% 88.24% 70.64% 31.15% 32 96.88% 90.69% 80.33% 35.04% 97.42% 94.84% 88.34% 54.29% % 37 96.38% 92.46% 88.24% 53.92% 97.65% 95.08% 91.59% 60.64% 42 98.25% 96.54% 92.58% 53.46 % 43 98.46% 95, 60% 89.21% 38.85% 46 98.77% 94.81% 84.23% 48.74% 47 94.60% 76.71% 52.94% 30, 18% siRNAs according to the present invention show greater than 94% efficiency at 1 nM for the most efficient sequences. The lowest dose tested is 0.016 nM for a 72% efficiency on degradation of mRNA encoding tyrosinase.

  EXAMPLE 3 Evaluation of a Possible Interferon-type Response Caused by the siRNAs According to the Invention It was verified that the siRNAs of the present invention did not induce an interferon type response. For this, the expression of the OAS-1 and IFIT-1 (interferon-inducible tetratricopeptide repeat domain) genes known to be induced by interferon (Wieland SF et al., Searching for Interferon-Induced Genes That Inhibit Hepatitis B Virus Replication in Transgenic Mouse Hepatocytes, J. of Virology 2003; 77: 1227-1236 Persengiev SP et al., Nonspecific, concentration-dependent stimulation and repression of mammalian expression by small interfering RNAs (siRNAs). RNA 2004; 10: 12 Bridge AJ et al., Induction of an interferon response by RNAi vectors in mammalian cells Nature Genet 2003; 34: 263-264 Scacheri PC et al., Short interfering RNAs can induce unexpected and divergent changes in the levels of Untargeted Proteins in Mammalian Cells, Proc Natl Acad Sci USA 2004, 101: 1892-1897 Sledz CA et al., Activation of the interferonsystem by short-interfering RNAs Nat Cell Biol., 2003; 9: 834- 9. Patzwahl R et al., Enhanced expression of interferon-regulated genes in the liver f patients with chronic hepatitis C virus infection: detection by suppression-subtractive hybridization. J. Virol. 2001; 75: 1332-1338) was measured by quantitative PCR (qPCR) according to the protocol described in the kit manual BLOCK-iTTM 'RNAi Stress Response Control Kit (Human) for interferon-mediated monitoring stress response to double-stranded RNA in human cells marketed by Invitrogen.

  Measurement of the level of expression of the hOAS-1 gene: The level of expression of the hOAS-1 gene has been reported to that of the GAPDH reference gene.

  Sample hOAS-1 / GAPDH Stdev dsRNA from SEQ. ID. n 1 - 1 nM 0.608 0.046 dsRNA from SEQ. ID. n 2 - 1 nM 0.668 0.292 dsRNA from SEQ. ID. n 4 - 1 nM 0, 608 0.070 dsRNA from SEQ. ID. n 13 - 1 nM 1,232 0,866 dsRNA from SEQ. ID. n 46 - 1 nM 0.911 0.221 dsRNA from SEQ. ID. n 1 - 50 nM 1,080 0,500 Control sequence at 1 nM 2,141 1,336 Positive control (1 kb long dsRNA) 24,905 6,431 dsRNA of SEQ. ID. n 1 - 0.1 nM 0.856 0.534 dsRNA from SEQ. ID. n 2 - 0.1 nM 0.866 0.300 dsRNA from SEQ. ID. n 4 - 0.1 nM 0.966 0.568 dsRNA from SEQ. ID. n 13 - 0.1 nM 1,238 0,451 dsRNA of SEQ. ID. n 46 - 0.1 nM 1.262 0.328 Control sequence at 50 nM 0.804 0.404 Control sequence at 0.1 nM 1.949 1.502 Result: Only the positive control (long dsRNA of 1 kb) induces a significant increase in gene expression Hoas-1. The sequences tested (SEQ ID Nos. 1, 2, 4, 13 and 46) do not significantly increase the expression of the hOAS-1 gene.

  Measurement of the expression level of the hIFIT-1 gene Stdev dsRNA threshold cycle sample of SEQ. ID. n 1 - 1 nM 36.15 0.20 dsRNA from SEQ. ID. n 2 - 1 nM 32.77 1.85 dsRNA from SEQ. ID. n 4 - 1 nM 35.98 0.88 dsRNA from SEQ. ID. n 13 - 1 nM 35.41 1.88 dsRNA from SEQ. ID. n 46 - 1 nM 34.42 3.24 dsRNA from SEQ. ID. n 1 - 50 nM 34.16 3.02 1 nM control sequence 31.30 1.98 Positive control (1 kb long dsRNA) 25.70 0.79 dsRNA from SEQ. ID. n 1 -0.1 nM 32.47 3.07 dsRNA from SEQ. ID. n 2 - 0.1 nM 35.38 1.41 dsRNA from SEQ. ID. n 4 - 0.1 nM 34.86 1.86 dsRNA from SEQ. ID. n 13 - 0.1 nM 33.22 2.33 dsRNA from SEQ. ID. n 46 - 0.1 nM 33.89 2.03 Control sequence at 50 nM 35.70 3.30 Control sequence at 0.1 nM 33.65 3.90 Result: only the positive control (long dsRNA of 1 kb) induces a significant expression of the hIFIT-1 gene detected at a threshold cycle value of approximately 25. The sequences tested (SEQ ID Nos. 1, 2, 4, 13 and 46) do not induce the expression of the gene. hIFIT-1 only for threshold cycle values of about 32-36 is a difference of 7-11 cycles corresponding to a 102-103 times smaller amount of messengers.

  These tests make it possible to conclude that the dsRNAs according to the invention do not induce an interferon type response.

Claims (16)

  A composition comprising at least one double-stranded RNA oligonucleotide of sequence chosen from SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48 associated with a particle cationic of a size less than or equal to 1 μm, with a zeta potential of between 10 and 80 mV.   2. Composition according to claim 1, characterized in that the oligonucleotide is modified by the addition to 2 'of a -OMethyl group.   3. Composition according to claim 1 or 2, characterized in that the double-stranded RNA oligonucleotide is present at a concentration of less than or equal to 100 M. 4. Composition according to any one of claims 1 to 3, characterized in that the cationic particle is selected from surfactant micelles, block polymer micelles, nonionic and cationic surfactant liposomes, niosomes, oleosomes, nanoemulsion particles, nanocapsules, organic particles or inorganic particles.   5. Composition according to any one of claims 1 to 4, characterized in that the cationic particle has a size less than or equal to 500 nm.   6. Composition according to any one of claims 1 to 5, characterized in that the cationic particle has a size less than or equal to 300 nm.   7. Composition according to any one of claims 1 to 6, characterized in that the cationic particle is a micelle of nonionic amphiphilic surfactants and cationic surfactants.   8. Composition according to any one of claims 1 to 6, characterized in that the block polymer micelle is selected from a micelle of cationic amphiphilic block polymer, a micelle of nonionic amphiphilic block polymer and cationic amphiphilic block polymer and a nonionic amphiphilic block polymer micelle and cationic surfactant.   9. Use of at least one double-stranded RNA oligonucleotide of sequence chosen from SEQ. ID. n 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 , 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48 to inhibit the expression of tyrosinase.   10. Use of at least one double-stranded RNA oligonucleotide of sequence chosen from SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48 to inhibit expression of tyrosinase, characterized in that said oligonucleotide is associated with a cationic particle having a size of less than or equal to 1 μm and a zeta potential of between 10 and 80 mV.   Use according to claim 9 or 10, as a skin whitening agent and / or as an anti-browning agent.   12. Use according to any one of claims 9 to 11, to prevent the formation and / or eliminate pigment spots, senescence spots and / or to lighten and / or whiten and / or uniformize the color of a skin browned.   SEQ. ID. n 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 , 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48, 14. Use of at least one double-stranded RNA oligonucleotide of sequence selected from SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48, for the preparation of a composition for depigmenting the skin, characterized in that said oligonicleotide is associated with a cationic particle having a size of less than or equal to 1 μm and a zeta potential of between 10 and 80 mV.   15. Use according to claim 14, characterized in that the composition is intended to treat idiopathic melasmas, hyperpigmentations associated with pregnancy or estrogen-progestin contraception, accidental hyperpigmentations, hyperpigmentations due to leukoderma, vitiligo.   16. Cosmetic process for whitening and / or lightening the complexion and / or uniformizing the color of a burnished skin comprising the topical application of at least one double-stranded RNA oligonucleotide of sequence chosen from SEQ. ID. n 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 13. Use of at least one double-stranded RNA oligonucleotide of the sequence chosen for the preparation of a composition for depigmenting the skin.   15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48.   17. A cosmetic process for whitening and / or lightening the complexion and / or uniformizing the color of a burnished skin comprising the topical application of at least one double-stranded RNA oligonucleotide of sequence chosen from SEQ. ID. n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 , 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 or 48, characterized in that said oligonicleotide is associated with a cationic particle having a size of less than or equal to 1 μm and a zeta potential of between 10 and 80 mV.   18. The method of claim 16 or 17, characterized in that said oligonucleotide is formulated in a topical composition at a concentration less than or equal to 100 p.M.   19. Method according to any one of claims 16 to 18, characterized in that the topical application is preceded by a pretreatment of the skin selected from a stripping, a chemical peel, a mechanical dermabrasion, the application of a or a plurality of solvents having a delipidating effect, the cleaning of the skin with a detergent foaming product 20. A method according to any one of claims 16 to 19, characterized in that the topical application is followed by an occlusive post-treatment.